This application is a 371 of PCT/CZ98/00046 filed Oct. 2, 1998.
The invention relates to a preparation for prevention and healing of inflammation affections having a radical scavenging capacity with a large biological spectrum coverage. The preparation is applicable to the prevention and healing of skin, hide, fascia and muscle surface injuries and wounds extending in various depths which are accompanied by a massive proliferation of radicals due to the oxygen reduction and by generation of reactive oxygen products.
Due to the effect of various types of radiation such as UV, gamma, X-ray and similar radiation as well as hyperoxia, xenobiotic, upon injury or as a result of certain diseases, the living organisms, may suffer from surface lesion affecting the skin, fascia, and muscle in different depth. Upon-any injury a massive proliferation of radicals occurs due to the oxygen reduction accompanied by generation of reactive oxygen compounds. The reason for the radical proliferation is a defect in coordination of redox enzymatic systems of the living tissue under injury as well as the activity of the leukocytes present. The reactive oxygen compounds, mostly of radical nature, have aggressive impact on biological systems and produce often irreparable changes, for example in reaction with lipids, proteins, or DNA and cause damage to physiological protective mechanisms preventing the biological systems from the adverse effect of reactive oxygen products. Simultaneously, enzymatic systems are activated, Which contributes to the generation of the reactive oxygen products.
The protective systems present physiologically in the living organisms are low-molecular weight compounds such as vitamins C, E, glutathione or macromolecular compounds such as enzymes, catalase, superoxide dismutase, glutathione, reductase, peroxidase or cyclooxygenases. In the event of a deep tissue damage the aforesaid compounds align with the phagocyte activity of leukocytes are not able to control the proliferation of free radicals to the extent which could ensure a satisfactory healing effect.
If a wound is attacked by bacteria, the reactive oxygen radicals may cause damage to the tissue but, on the other hand, they are not enough to stop the bacteria growth, which results in the degradation of the tissue macromolecules and in the accompanying penetration of leukocytes, which is the major reason for a pyogenic process. Subsequently, even if the production of bacterial flora is controlled, such radicals cause an excessive cytokine production, which promotes the growth of fibroblasts and, due to copious granulation, the epithelization is going down, which again results in a delayed healing process.
The conventional medical treatment mostly concentrates on the inflammation stage (using bacteria eliminating antibiotics, granulation medicaments, including prostaglandins) and is mostly insufficient in the epithelization stage, for example, a treatment of the wound may occur, however, granulations are poor or granulations are copious but the wound fails to come to epithelization.
The object of the invention of a preparation for prevention and healing of inflammation affections is to eliminate, to a considerable extent, the above inconvenience. The preparation comprises 0.1 to 99.9% by weight, derivatives of sterically hindered amines selected from the group consisting of:
soluble polymers or copolymers prepared by radical polymerization in the presence of 0.01 to 10% by weight of initiators in polymerization mixture comprising individually or in combination an aliphatic amine monomer of the general formula (A): 
where RA to RF are: alkyl C1-C4,xe2x80x94(CH2)nxe2x80x94 and n=3, 4, 5, polymerizable vinyl group in various combination and representation, R6 is alkyl C1-C4, individually or in any combination, H, OH or oxygen radical obtained by additional oxidation; soluble polymers or copolymers prepared by radical polymerization in the presence of 0.01 to 10% by weight of initiators, from monomers of cyclic sterically hindered amines of general formula (B) 
where R1 to R4 are: alkyl C1-C4, xe2x80x94(CH2)nxe2x80x94 and n=3, 4, 5, R6 is alkyl C1-C4, individually or in any combination, H, OH or oxygen radical obtained by additional oxidation and W is selected from the group including xe2x80x94CH(X)xe2x80x94 a xe2x80x94CH(X)CH2xe2x80x94 where X is: 
xe2x80x94N(X)xe2x80x94 and xe2x80x94N(X)CH2xe2x80x94 where X is: 
xe2x80x94Oxe2x80x94, xe2x80x94OCH2xe2x80x94, and R1 to R4 is a radical having one polymerizable vinyl group; soluble polymers prepared by polycondensation of difunctional sterically hindered amimes of the general formula (F) 
where RA to RF are: alkyl C1 to C4, xe2x80x94(CH2)nxe2x80x94 and n=3, 4, 5, hydroxyalkyl, aminoalkyl, carboxyalkyl (halogenide, activated ester, azide), isocyanatoalkyl in various combination and representation, R6 is alkyl C1-C4, individually or in combination H, OH or oxygen radical obtained by additional oxidation;
soluble polymers or copolymers prepared by polycondensation of difunctional cyclic sterically hindered amines of the general formula ((G) 
where R1 to R4 are: alkyl C1-C4, xe2x80x94(CH2)nxe2x80x94 and n=3, 4, 5, hydroxyalkyl, aminoalkyl, carboxyalkyl (halogenide, activated ester, azide), isocyanatoalkyl, R6 is alkyl C1-C4, individually or in combination H, OH or oxygen radical obtained by additional oxidation and W is selected from the group xe2x80x94Oxe2x80x94, xe2x80x94OCH2xe2x80x94, xe2x80x94NHxe2x80x94, xe2x80x94NHCH2xe2x80x94 where R1-R4 is hydroxyalkyl, aminoalkyl, carboxyalkyl (halogenide, activated ester, azide), isocyanatoalkyl) xe2x80x94CH(X)xe2x80x94, xe2x80x94CH(X)CH2xe2x80x94, where X is: 
n is 1 to 10, m is 2 to 10
where A, B are xe2x80x94OH, xe2x80x94NH2, xe2x80x94COOH;
soluble copolymers prepared by polycondensation of difunctional cyclic, sterically hindered amines of the general formula (G) and monofunctional cyclic sterically hindered amines in an amount of 0.1 to 15%. by weight, based on the total polymerization mixture of the general formula (H) 
where R1 to R4 are: alkyl C1-C4, xe2x80x94(CH2)n and n=4 or 5, hydroxyalkyl, aminoalkyl, carbokyalkyl, ore their reactive derivatives, R6 is alkyl C1-C4, individually or in combination H, OH or oxygen radical obtained by additional oxidation and W is selected from the group consisting of: xe2x80x94Oxe2x80x94, OCH2xe2x80x94, xe2x80x94NHxe2x80x94, xe2x80x94NHCH2xe2x80x94, where R1 to R4 is hydroxyalkyl, aminoalkyl, carboxyalkyl, CH(X)xe2x80x94 a CH(X)CH2xe2x80x94 where X: xe2x80x94COOH (halogenide, activated ester, mixed anhydride, azide), xe2x80x94NCO, 
where n is 2-10, m is 1-10;
derivatives of sterically hindered cyclic amines of general formula 
where R1 to R4 are: alkyl C1-C4, xe2x80x94(CH2)nxe2x80x94 and n=3 to 5, hydroxyalkyl, aminoalkyl, carboxyalkyl, in combination and any representation R6 is alkyl C1 to C4, H, OH or oxygen radical in any representation and W is represented by following groups: xe2x80x94Oxe2x80x94, xe2x80x94OCH2xe2x80x94, xe2x80x94NHxe2x80x94, xe2x80x94NHCH2xe2x80x94, xe2x80x94CH(X)xe2x80x94. xe2x80x94CH(X)CH2xe2x80x94, where X=
and R1, R2 are alkyl C1-C10;
polymers, copolymers, natural compounds comprising free reactive groups xe2x80x94OH, xe2x80x94NH2, xe2x80x94COOH, xe2x80x94CHO, oxiran, selected from the group consisting of poly(vinyl alcohol), cellulose, (2-hydroxyethyl)cellulose, (carboxymethyl)celullose, agar derivatives, polymers obtained by condensation, using dihydroxyalkane derivatives, oligomers and polymers of ethylene glycol or propylene glycol, natural o synthetic polymers, comprising a free carboxyl group, amino group or aldehyde group, prepared by additional functionalization of polymers or natural compounds by polymer-analogous reaction with a suitable sterically hindered amine selected from the group comprising:
4-X-1-R6-2,2,6,6-tetramethyl-piperidine 
where X is xe2x80x94NH2,i xe2x80x94OH, -halogen, xe2x80x94NCO, xe2x80x94COOH (halogenide, activated ester, mixed anhydride, azide), xe2x80x94CH2Br and R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation;
(n-X-alkyl)-1-R6-(2,2,6,6-tetramethyl-piperidin-4-yl)amine of the general formula 
where n=1 to 10, X is halogen, xe2x80x94OH, xe2x80x94NH2, xe2x80x94COOH, (halogenide, mixed anhydride, activated ester, azide), R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation,
1-R6-(2,2,6,6-tetramethylpiperidin-4-yl)-(n+1)-X-alkanoate of general formula 
where n=1-10, X is xe2x80x94OH, xe2x80x94NCO, xe2x80x94COOH (halogenide, activated ester, mixed anhydride, azide), xe2x80x94NCO, R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation;
4-(n-X-alkyl)-1-R6-2,2,6,6-tetramethylpiperidine of the general formula: 
where n=1-10, X is xe2x80x94NH2, -halogen, xe2x80x94OH, xe2x80x94NCO, xe2x80x94COOH (halogenide, activated ester, mixed anhydride, azide), R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation;
3-X-1-R6-2,2,5,5-tetramethylpyrrolidine of general formula: 
where X is xe2x80x94NH2, xe2x80x94OH, xe2x80x94NCO, xe2x80x94COOH, (halogenide, activated ester, mixed anhydride, azide), xe2x80x94CH2Br, R6 is alkyl C1 to C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation;
1-R6-(2,2,5,54tetramethyl-pyrrolidin-3-yl)(n+1)-X-alkanoate of the general formula: 
where n is 1 to 10, X is xe2x80x94NH2, xe2x80x94OH, xe2x80x94NCO, xe2x80x94COOH (halogenide, activated ester, mixed anhydride, azide), R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation;
1-R6-2,2,5,5-tetramethyl-2,5-dihydropyrrole-3-carboxylic acid 
where A is xe2x80x94COOH, (chloride, mixed anhydride, activated ester, azide), glycidyl ester, R6 is alkyl C1-C4, individually or in combination, H, OH or oxygen radical obtained by additional oxidation.
Further developments and improvements of the invention are demonstrated by reference to various compounds which are subject to dependent claims and the preparation and effects of which are demonstrated in the accompanying examples. The invention is based on a new finding that amines with sterically hindered amino groups are able to substantially liquidate reactive oxygen derivatives and thus to enable accelerated healing of damaged tissues. The mechanism of their effect on the living organism has not been described, however, the chemism of the elimination of the oxygen reactive radicals may be similar to that described for the polymeric systems. The presence of any of the oxidation states of such amines (hydroxylamine or nitroxide) accelerates the healing of damaged tissues since, due to the recombination with free radicals or oxidation agents, such as hydrogen peroxide, organic peroxy radicals, hyperoxides, etc. formed in the living tissue, a considerably larger spectrum of compounds for their liquidation is available.
The compounds contained in the preparation according to the invention include soluble or crosslinked polymers, which are formed from polymerizable sterically hindered amines or copolymers, wherein, besides a sequence formed from a polymerizable amine, any suitable monomer may be incorporated, advantageously a hydrophilic one. The accessibility of the amino group is from the steric point of view substantially restricted. The stabile free radicals derived from that group of compounds are not able to initiate a radical polymerization. The sterically hindered amines may survive the polymerization in their original form or, i.e., as amine or, as the case may be, in a higher oxidation state i.e. as hydroxylamine or nitroxide.
This type of amines or their derivatives preferably react with oxygen and its reduced derivatives such as superoxide, hydroxyl radical, hydrogen peroxide, alkyl peroxides, alkyl hyperoxides, etc. and protect the tissue from destructive oxidation. The hydrophilic polymers are suitable for applications in medicine in the form of gels, foils, therapeutic.contact lenses, powders, etc. In a proper configuration such as foam, sponge, etc. The aforesaid polymeric systems may simultaneously remove water from the injured tissues.
The same effect results in production of polymers prepared by condensation of di- or polyfunctional alcohol, amines, aminoalcohol with reactive derivatives of di- and polyfunctional acid derivatives such as chlorides, activated esters, mixed anhydrides, or bifunctional isocyanates, upon formation of polyesters, polyamides, urethanes or their combination compounds. In such polymers formed by condensation, it is of course assumed that they contain a built-in sterically hindered amine provided with suitable reactive groups. Such polymers prepared by condensation may be prepared, dependent on the polymerization condition, in a soluble or crosslinked form.
In connection with the new preparation, the antibacterial effect of quaternary ammonium salts may be used and in this case the therapeutic effect of the polymeric derivatives of the sterically hindered amines and their oxidation derivatives may be combined with the therapeutic effect of the polymeric derivatives of quaternary ammonium salts in order to achieve substantially prolonged period, which may be necessary for the application of polymeric systems, without causing a contamination. The enhanced antibacterial effect, for example with a denture prosthesis may be secured by the presence of bonded quaternary ammonium salts which may be prepared by copolymerization of the above specified polymerization mixtures with the polymerizable quaternary salts or precursors thereof and their subsequent quaternization or by condensation of substituted amines in such a manner that the quaternization may be carried out subsequently.
The method of preparation of copolymers consists in the polymerization of a polymerization mixture including some of the above described vinyl monomers or a combination thereof, a polymerizable sterically hindered amine or a mixture thereof or, as the case may be, a hydroxylamine derivative and, if necessary, a crosslinking agent and initiator under formation of a soluble or insoluble polymer powder or a required article formed in the polymerization mold such as foil, lens, etc. In preparation of hydrophilic gel articles, the process may be managed in such a way that in the first stage powdered hydrophilic polymers with a sterically hindered amine and amino group in various oxidation state represented by amine, hydroxylamine, or nitroxide radical or various types of derivatives of sterically hindered secondary amines in the form of powdered polymers having various oxidation state which may be combined, are prepared whereas in the second stage, the desired gel is prepared from such mixtures.
The advantage of the gel copolymer structures is that the gel may perfectly cover the lesion to be cured. Various promoting medicament forms may also be added to such gels prepared by the above process.
The polymers prepared by polycondensation are polyesters, polyamides, polyurethanes, or their mixtures. The polycondensation is carried out either with a bifunctional sterically hindered secondary amine or in the presence of other bifunctional monomers. The preparation of such polymers is governed by common rules of preparation of polymers by polycondensation.
In both soluble and insoluble polymers formed by condensation, quaternary ammonium salts may be incorporated therein by means of suitable derivatives. The precursors must be additionally quaternized.